Fluorescent-Antibody Techniques for the Identification of Group D Streptococci: Direct Staining Method1

Pavlova, Maria T.; Beauvais, Elizabeth; Brezenski, Francis T.; Litsky, Warren

doi:10.1128/aem.23.3.571-577.1972

Cited by 14 publications

(3 citation statements)

References 10 publications

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“…Type-specific carbohydrates, on the other hand, are clearly exposed on the surface of both group D and group B streptococci (4). The selected antisera used to prepare group-specific anti-B and anti-D reagent staphylococci in our investigations might also detect such typespecific antigens, thereby representing polytype reagents as has been discussed by others using fluorescent techniques (7,17). The group-specific antisera used in the present investigations were selected from among those not showing cross-reactions when coated on staphylococci and tested against a panel of streptococcal strains representing group A, B, C, D, and G. Most antisera screened could not be used because of such cross-reactions.…”

Section: Discussionmentioning

confidence: 88%

“…Some of the other sera showed cross-reactivities. The variability among different commercial and noncommercial antisera has previously been pointed out in immunofluorescence studies by Franek et al 7and Pavlova et al (17).…”

Section: Kronvall In Press)mentioning

confidence: 84%

“…The group determination of streptococci is most commonly performed by using various precipitation techniques with group-specific antisera and extracts of streptococcal group antigens (3,8,14,16,18,22) and also by immunofluorescent methods (7,17). Direct agglutination of untreated streptococci by group-specific antisera does not work regularly for two reasons: first, group-specific antigens are usually not exposed, and secondly, many strains show saline agglutinability (15,19).…”

Section: Kronvall In Press)mentioning

confidence: 99%

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New Method for the Serological Grouping of Streptococci with Specific Antibodies Adsorbed to Protein A-Containing Staphylococci

et al. 1973

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A coagglutinating reagent has recently been developed and utilized in capsule typing of pneumococci. The reagent consists of stabilized staphylococci coated with specific antibody via the gamma globulin Fc-protein A reaction thereby orienting the antibody molecules with the antigen-reactive Fab parts outwards. Reagent staphylococci coated with antibodies directed against streptococcal group-specific antigens have been used in the present investigations. Overnight cultures of streptococci were treated with trypsin and then directly tested for coagglutination of selected reagent staphylococci on glass slides. In all, 179 strains of streptococci were analyzed by the method described. The results were compared with grouping by using Lancefield extracts in counter-immunoelectrophoresis and were shown to agree completely. The coagglutination method described proved to be accurate, rapid, and simple.

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Section: Discussionmentioning

confidence: 88%

Section: Kronvall In Press)mentioning

confidence: 84%

Section: Kronvall In Press)mentioning

confidence: 99%

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New Method for the Serological Grouping of Streptococci with Specific Antibodies Adsorbed to Protein A-Containing Staphylococci

et al. 1973

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Viable microorganism detection by induced fluorescence

Snyder

Greenberg

1984

Biotech & Bioengineering

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A methodology for rapid detection and identification of microorganisms has long been of concern to the medical, pharmaceutical, and food processing fields, among others. Because of this sustaining interest the area has remained active and has seen significant advances over the classical time-consuming protocols of standard plate counting membrane filtration, or the multiple-tube fermentation procedures. However, even today these techniques can require as much as one to three days; hence, continued research is still indicated before a rapid, generalized microbial characterization scheme is to be realized.Various earlier developments have focussed upon techniques in immunofluorescence, 1-6 enzyme amplificat i~n , "~ light detection and ranging (LIDAR),9 mass spectrometry, l o bacteriophage lysis, ' ] -13 computer-assisted probabilistic methods, 14-18 the recently developing field of gel ferrography,'9120 and the double fluorescent DNA staining method coupled to flow cytometry.21 Each of these techniques is limited by disadvantages such as laborious and time-consuming laboratory preparation and sample handling, long observation times, and nonspecificity with respect to bacterial characterization and subsequent identification.The present investigation, which we believe to offer the possibilities of speed, sensitivity, and specie identification to the process of microorganism detection, is embraced in the following equation:This method combines several attractive experimental features. These include (1) exploitation of the particular enzymatic/metabolic machinery of the cell by an added substrate in which a predetermined reaction takes place;(2) the product fluorescence which can be used to indicate a positive cellular response; and (3) rapid data collection and analysis under 30 min. Equation (1) presents various modes of biochemical strategy in order to uniquely characterize a given bacterium or small group of bacteria. The technique pinpoints the identity of a particular enzyme that is indigenous to the bacteria themselves through several experimental avenues. In one instance, for the same bacterial enzyme a series of chemically similar substrates (e.g., esters) can be utilized to generate unique reaction rates and, thus, associated product fluorescence. Another scheme involves sequentially using a series of substrates in order to trigger the various potential enzymatic reactions that can occur within the bacterium. The present experimental format exploits the former procedure.The first-order nature of the experimental fluorescence producing chemistry in this work suggests the MichaelisMenten data analysis approach as most attractive. Precedence for applying Michaelis-Menten kinetics to enzymatic processes in vivo (i.e., reactions occurring in the intact, living cell) has been previously documented in the l i t e r a t~r e~~.~~ although the theory, strictly speaking, was initially developed for enzymatic processes occurring in vitro .The specific reaction examined was the hydrolysis of the substrate diacetylfluore...

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